The roboocyte: automated electrophysiology based on Xenopus oocytes.

Automated electrophysiological assays are of great importance for modern drug discovery, and various approaches have been developed into practical devices. Here, we describe the automation of two-electrode voltage-clamp (TEVC) recording from Xenopus oocytes using the Roboocyte automated workstation, jointly developed by Multi Channel Systems and Bayer Technology Services. We briefly discuss the technology, including its advantages and limitations relative to patch clamp and other TEVC systems. We provide a step-by-step description of typical operating procedures and show that the Roboocyte represents a practical and highly effective way to perform automated electrophysiology in an industrial setting.

[1]  D. Melton,et al.  Gene transfer in amphibian eggs and oocytes. , 1981, Annual review of genetics.

[2]  D. Bertrand,et al.  Electrophysiology of Neuronal Nicotinic Acetylcholine Receptors Expressed in Xenopus Oocytes following Nuclear Injection of Genes or cDNAs , 1991 .

[3]  B. Sakmann,et al.  Rat brain serotonin receptors in Xenopus oocytes are coupled by intracellular calcium to endogenous channels. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Christine Leisgen,et al.  Automated higher-throughput compound screening on ion channel targets based on the Xenopus laevis oocyte expression system. , 2004, Assay and drug development technologies.

[5]  R. Miledi,et al.  Messenger RNA from human brain induces drug- and voltage-operated channels in Xenopus oocytes , 1984, Nature.

[6]  B. Sakmann,et al.  Role of acetylcholine receptor subunits in gating of the channel , 1985, Nature.

[7]  D. Bertrand,et al.  Neuronal Nicotinic Acetylcholine Receptors from Drosophila , 2000 .

[8]  Michael Fejtl,et al.  The roboocyte: automated cDNA/mRNA injection and subsequent TEVC recording on Xenopus oocytes in 96-well microtiter plates. , 2003, Receptors & channels.

[9]  Bert Sakmann,et al.  Molecular distinction between fetal and adult forms of muscle acetylcholine receptor , 1986, Nature.

[10]  R. Miledi,et al.  Translation of exogenous messenger RNA coding for nicotinic acetylcholine receptors produces functional receptors in Xenopus oocytes , 1982, Proceedings of the Royal Society of London. Series B. Biological Sciences.

[11]  J. Schroeder Heterologous Expression and Functional Analysis of Higher Plant Transport Proteins in Xenopus Oocytes , 1994 .

[12]  J. Gurdon,et al.  Use of Frog Eggs and Oocytes for the Study of Messenger RNA and its Translation in Living Cells , 1971, Nature.

[13]  D. Sharon,et al.  Positive and Negative Coupling of the Metabotropic Glutamate Receptors to a G Protein–activated K+ Channel, GIRK, in Xenopus Oocytes , 1997, The Journal of general physiology.

[14]  A. Hodgkin,et al.  Measurement of current‐voltage relations in the membrane of the giant axon of Loligo , 1952, The Journal of physiology.